LOCUS       sequence-388597-        6831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SLC7A11 with an N-terminal FLAG tag in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6831)
  AUTHORS   Parker JL, Deme JC, Kolokouris D, Kuteyi G, Biggin PC, Lea SM, 
            Newstead S
  TITLE     Molecular basis for redox control by the human cystine/glutamate 
            antiporter system xc().
  JOURNAL   Nat Commun. 2021 Dec 8;12(1):7147. doi: 10.1038/s41467-021-27414-1.
  PUBMED    34880232
REFERENCE   2  (bases 1 to 6831)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2021-12-8"; volume: "12(1):7147. doi"; pages: " 
            10.1038/s41467-021-27414-1"
COMMENT     Sequence Label: pLexM-SLC7A11
FEATURES             Location/Qualifiers
     source          1..6831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..663
                     /label=chicken beta-actin promoter
     intron          664..1680
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1604..1626
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1688..1707
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1774..1783
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1783..1806
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3319..3339
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3346..4056
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             4057..4080
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(4186..4205)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4251..4306
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4305..4324)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4667..4683)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4667..4683)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4680..4702)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4691..4707
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4715..4721,4722..4739,4740..4745))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4760..4781
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4910..4927)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5081..5669)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5161..5180)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5840..6631,6632..6700))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6463..6482
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6701..6805)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gcgccrggsg gggsggggsg gggsgrgggg
      541 sggggsgggg sgrggcggag aggkgcggcg gcagccaatc agagcggcgc gctccgaaag
      601 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
      661 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
      721 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
      781 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
      841 gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg
      901 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag
      961 cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc
     1021 gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt
     1081 gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg
     1141 cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg
     1201 cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg
     1261 gcggggccgc ctcgggccgg ggrgggctcg ggggaggggc gcggcggccc ccggagcgcc
     1321 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
     1381 gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc
     1441 accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg
     1501 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt
     1561 ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt
     1621 gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag
     1681 ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attgcggccg
     1741 tctcaggccg aattcaacgt tggtaccgag ctcgccacca tggactacaa agacgatgac
     1801 gacaaggtca gaaagcctgt tgtgtccacc atctccaaag gaggttacct gcagggaaat
     1861 gttaacggga ggctgccttc cctgggcaac aaggagccac ctgggcagga gaaagtgcag
     1921 ctgaagagga aagtcacttt actgagggga gtctccatta tcattggcac catcattgga
     1981 gcaggaatct tcatctctcc taagggcgtg ctccagaaca cgggcagcgt gggcatgtct
     2041 ctgaccatct ggacggtgtg tggggtcctg tcactatttg gagctttgtc ttatgctgaa
     2101 ttgggaacaa ctataaagaa atctggaggt cattacacat atattttgga agtctttggt
     2161 ccattaccag cttttgtacg agtctgggtg gaactcctca taatacgccc tgcagctact
     2221 gctgtgatat ccctggcatt tggacgctac attctggaac cattttttat tcaatgtgaa
     2281 atccctgaac ttgcgatcaa gctcattaca gctgtgggca taactgtagt gatggtccta
     2341 aatagcatga gtgtcagctg gagcgcccgg atccagattt tcttaacctt ttgcaagctc
     2401 acagcaattc tgataattat agtccctgga gttatgcagc taattaaagg tcaaacgcag
     2461 aactttaaag acgccttttc aggaagagat tcaagtatta cgcggttgcc actggctttt
     2521 tattatggaa tgtatgcata tgctggctgg ttttacctca actttgttac tgaagaagta
     2581 gaaaaccctg aaaaaaccat tccccttgca atatgtatat ccatggccat tgtcaccatt
     2641 ggctatgtgc tgacaaatgt ggcctacttt acgaccatta atgctgagga gctgctgctt
     2701 tcaaatgcag tggcagtgac cttttctgag cggctactgg gaaatttctc attagcagtt
     2761 ccgatctttg ttgccctctc ctgctttggc tccatgaacg gtggtgtgtt tgctgtctcc
     2821 aggttattct atgttgcgtc tcgagagggt caccttccag aaatcctctc catgattcat
     2881 gtccgcaagc acactcctct accagctgtt attgttttgc accctttgac aatgataatg
     2941 ctcttctctg gagacctcga cagtcttttg aatttcctca gttttgccag gtggcttttt
     3001 attgggctgg cagttgctgg gctgatttat cttcgataca aatgcccaga tatgcatcgt
     3061 cctttcaagg tgccactgtt catcccagct ttgttttcct tcacatgcct cttcatggtt
     3121 gccctttccc tctattcgga cccatttagt acagggattg gcttcgtcat cactctgact
     3181 ggagtccctg cgtattatct ctttattata tgggacaaga aacccaggtg gtttagaata
     3241 atgtcagaga aaataaccag aacattacaa ataatactgg aagttgtacc agaagaagat
     3301 aagttatgag tcgaggggga aaatttatat tttcaaggtc aattttctaa aggtgaagaa
     3361 ttattcactg gcgttgtccc aattttggtt gaattagatg gtgatgttaa tggtcacaaa
     3421 ttttctgtct ccggtgaagg tgaaggtgac gctacttacg gtaaattgac cttaaaattt
     3481 atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac tttcggttat
     3541 ggtgttcaat gttttgcgag atacccagat catatgaaac aacatgactt tttcaagtct
     3601 gccatgccag aaggttatgt tcaagaaaga actatttttt tcaaagatga cggtaactac
     3661 aagaccagag ctgaagtcaa gtttgaaggt gataccttag ttaatagaat cgaattaaaa
     3721 ggtattgatt ttaaagaaga tggtaacatt ttaggtcaca aattggaata caactataac
     3781 tctcacaatg tttacatcat ggctgacaaa caaaagaatg gtatcaaagt taacttcaaa
     3841 attagacaca acattgaaga tggttctgtt caattagctg accattatca acaaaatact
     3901 ccaattggtg atggtccagt cttgttacca gacaaccatt acttatccac tcaatctgcc
     3961 ttatccaaag atccaaacga aaagagagac cacatggtct tgttagaatt tgttactgct
     4021 gctggtatta cccatggtat ggatgaattg tacaaacacc accaccatca ccatcatcat
     4081 taagtcgaga ctagtatcgc gataattcac tcctcaggtg caggctgcct atcagaaggt
     4141 ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc
     4201 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa
     4261 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     4321 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata
     4381 tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa
     4441 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt
     4501 ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt
     4561 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct
     4621 cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc
     4681 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4741 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4801 ccgctttcca gtcgggaaac ctgtcgtgcc agcggatcga tccgctgcat taatgaatcg
     4861 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4921 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4981 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5041 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5101 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5161 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5221 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5281 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5341 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5401 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5461 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5521 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5581 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5641 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5701 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5761 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5821 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5881 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5941 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6001 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6061 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6121 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6181 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6241 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6301 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6361 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6421 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6481 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6541 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6601 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6661 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6721 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6781 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct g
//
